Nanomotors convert chemical or other energy sources into motion, exerting forces that can move an external cargo. They are an important part of nanotechnology. The overall goal of this project is to develop a biochemical nanomotor that moves along a linear polymer. Such a device will be used as a module within a powered nanodevice. It will be controlled and directed so as to deliver cargo along specified tracks to specific addresses. To that end we will use the ATP-dependent protein ClpX, which engages, unfolds and moves proteins. In its native bacterial context ClpX functions to feed protein substrates into a protease. We will adapt it to move instead along DNA, thereby using ClpX as an ATP-powered winch with DNA as a cable. In furtherance of these goals we will perform the following experiments. Specific Aim 1: Construct chimeric objects consisting of (1) a protein or peptide that engages ClpX and initiates translocation, linked to (2) a DNA along which ClpX subsequently continues to move. Specific Aim 2: Develop and apply assays to assess the powered translocation of ClpX along DNA. Specific Aim 3: Using these assays, modify the properties of DNA to optimize translocation velocity and power. Use mutagenesis of E. coli ClpX and ClpX sourced from diverse bacterial species to improve the desired properties of the motor. PUBLIC HEALTH RELEVANCE: oject Narrative Nanomotors convert chemical or other energy sources into motion, exerting forces that can move an external cargo. They are an important part of nanotechnology. We will adapt the bacterial protein ClpX, which engages, unfolds and moves proteins, developing it to function as a novel biochemical nanomotor that moves along DNA.